Correlative biomarkers of IL-6 blockade combined with checkpoint inhibition

这一建议是基于对接受PD-1抗体治疗的黑色素瘤患者的血清进行分析的。 Nivolumab(Nivo)显示高水平的IL-6和IL-6样C-反应诱导的急性时相反应物(APP)。 反应蛋白(CRP)与生存不良有关。我们假设C反应蛋白和其他APP诱导 IL-6与黑色素瘤对检查点抑制的抵抗有关，并具有免疫抑制作用。 我们的初步数据显示，CRP水平与低应答率相关，这一点得到了支持 NIVO或ipilimumab(IPI)的短期生存期。IL-6促进CRP和其他APP的合成 肝脏及其水平也与低应答率、短生存期和治疗耐药有关 接受Nivo、IPI或联合免疫检查点阻断(ICB)的患者。白介素6水平低的患者 基线或ICB治疗后有较高的有效率和较长的生存期。克服免疫力 观察到CRP抑制和逆转与高IL-6相关的ICB耐药，我们将添加IL-6 转移性黑色素瘤患者中抗ipi和nivo的受体阻断抗体tocilizumab。我们最重要的是 假设是基线血清中IL-6和CRP水平以及血清、PBMC和肿瘤中其他生物标志物的水平 将在IPI/Nivo与tocilizumab的II期试验中预测疗效和毒性，IL-6阻断将 提高联合ICB的疗效，降低其毒性。这些生物标志物将提供对 ICB的宿主/肿瘤介质，并帮助指导患者进行最有效的免疫治疗 IL-6/CRP和其他生物标志物的基线和治疗中水平。试验的共同主要终点包括 自IL-6阻断以来最好的总体应答率和毒性也被证明可以减少与免疫相关的 免疫治疗的不良事件。SIMON两阶段设计试验将包括67名患者，这些患者将接受 三重组合，并将得到BMS的支持。在这项建议中，我们希望支持相关的 基于第二阶段试验的标记物研究。将进行血清、肿瘤和外周血的分析 建立疗效和毒性的生物标记物，并帮助了解联合耐药的基础 ICB在黑色素瘤中基于三个特定的目的：1.确定基线水平或治疗中的变化 在血清质谱仪-MALDI-TOF蛋白特征和急性期反应物和细胞因子 使用IL-6受体阻断抗体的IPI/NIVO II期试验与反应和毒性相关 Tocilizumab；2.确定基线或治疗中外周血和肿瘤T细胞是否发生变化 用多参数流式细胞术、抗体序列和等光法检测外周血细胞表型和功能 血树突状细胞表型和功能与肿瘤I、II类表达、PD-L1水平和RNA序列 在使用tocilizumab的IPI/Nivo的II期研究中，签名与反应和毒性有关；和3. 结合AIMS 1和AIMS 2的血清、外周血和肿瘤生物标记物来评估是否合并 在使用tocilizumab的IPI/Nivo的II期研究中，签名与反应和毒性有关。

This proposal is based on an analysis of serum from melanoma patients treated with the PD-1 antibody nivolumab (NIVO) showing that high levels of IL-6 and acute phase reactants (APP) induced by IL-6 like C- reactive protein (CRP) are associated with poor survival. We hypothesize that CRP and other APP induced by IL-6 are associated with resistance to checkpoint inhibition in melanoma, and are immunosuppressive. This is supported by our preliminary data showing that CRP levels are associated with low response rates and short survival with NIVO or ipilimumab (IPI). IL-6 promotes the synthesis of CRP and other APPs from the liver, and its levels are also associated with low response rates, short survival and therapeutic resistance in patients receiving NIVO, IPI or combination immune checkpoint blockade (ICB). Patients with low IL-6 at baseline or after treatment with ICB have high response rates and long survivals. To overcome the immune suppression observed with CRP and reverse ICB resistance associated with high IL-6, we will add the IL-6 receptor blocking antibody tocilizumab to IPI and NIVO in patients with metastatic melanoma. Our overarching hypothesis is that levels of IL-6 and CRP in baseline serum and other biomarkers in serum, PBMCs and tumor will predict efficacy and toxicity in a phase II trial of IPI/NIVO with tocilizumab and that IL-6 blockade will augment the efficacy of, and reduce toxicity from combined ICB. These biomarkers will provide insight into host/tumor mediators of ICB and help direct patients to the most effective immunotherapy based on their baseline and on-treatment levels of IL-6/CRP and other biomarkers. Co-primary endpoints of the trial include best overall response rate and toxicity since IL-6 blockade has also been shown to reduce immune-related adverse events with immunotherapy. A Simon two-stage design trial will include 67 patients that will receive the triple combination and will be supported by BMS. In this proposal, we wish to support the correlative marker studies based on that phase II trial. Analyses of the serum, tumor and peripheral blood will be carried out to establish biomarkers of efficacy and toxicity and help understand the basis for resistance to combination ICB in melanoma based on three specific aims: 1. To determine if baseline levels, or on-treatment changes in a serum mass spectrometry-MALDI-TOF protein signature and acute phase reactants and cytokines are associated with response and toxicity in a phase II trial of IPI/NIVO with the IL-6 receptor blocking antibody tocilizumab; 2. To determine if baseline or on-treatment changes in peripheral blood and tumor T cell phenotype and function assessed by multi-parameter flow cytometry, Ab-Seq and IsoLight assays, peripheral blood dendritic cell phenotype and function and tumor class I and II expression, PD-L1 levels and RNA seq signatures are associated with response and toxicity in a phase II study of IPI/NIVO with tocilizumab; and 3. To integrate the serum, peripheral blood and tumor biomarkers of aims 1 and 2 to assess if a consolidated signature is associated with response and toxicity in a phase II study of IPI/NIVO with tocilizumab.